Modified plants and methods of detecting pathogenic disease

Abstract

Genetically modified plants and methods of detecting diseases by a relevant color change in the genetically modified plant compared to a non-genetically modified plant are provided. Also disclosed is a system for remote detection of pathogens on crops and a methods for treating crops under a pathogenic attack.

The invention claimed is: 
     
       1. A modified plant cell comprising a cisgenic construct, said construct comprising:
 a promoter comprising a pathogen inducible regulatory element, wherein said promoter comprises a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46 and SEQ ID NO: 47, operably linked to a nucleic acid sequence encoding a signaling moiety, wherein said promoter and said nucleic acid sequence encoding the signaling moiety are both endogenous to said plant cell but are not operably linked to one another in the native plant cell. 
 
     
     
       2. The modified plant cell of  claim 1  wherein the cisgenic construct is generated by inserting either the promoter or the nucleic acid sequence encoding a signaling moiety into a genomic location that results in the operable linkage of the promoter to the nucleic acid sequence encoding a signaling moiety. 
     
     
       3. The modified plant cell of  claim 1  wherein said cisgenic construct is inserted into a novel genomic site of said plant cell. 
     
     
       4. A plant or plant part comprising a plurality of the modified plant cells according to  claim 1 . 
     
     
       5. The plant or plant part of  claim 4 , wherein said signaling moiety is an anthocyanin pathway factor. 
     
     
       6. The plant or plant part of  claim 5 , wherein said anthocyanin pathway factor is
 i) a transcription factor that enhances production of anthocyanins; or 
 ii) a rate limiting anthocyanin pathway enzyme. 
 
     
     
       7. The plant or plant part of  claim 5 , wherein said anthocyanin pathway factor is a transcription factor that enhances production of anthocyanins, said transcription factor being
 i) selected from the group consisting of C1, R, P11, and B1, or alleles of these genes; or 
 ii) a nucleic acid encoding an amino acid sequence selected from the group consisting of anthocyanin1 transcription factor (ANT1; SEQ ID NO: 34), Zm00001d044975 (SEQ ID NO: 27, Zm00001d026147 (SEQ ID NO: 28), Zm00001d000236 (SEQ ID NO: 29), Zm00001d028842 (SEQ ID NO: 30), Zm00001d008695 (SEQ ID NO: 31), Zm00001d037118 (SEQ ID NO: 33), Zm00001d019170 (SEQ ID NO: 33), and Glyma09g36990 (SEQ ID NO: 35). 
 
     
     
       8. The plant or plant part of  claim 5  wherein said plant or plant part is
 a corn plant wherein a plurality of cells of the corn plant comprise said promoter operably linked to said nucleic acid sequence encoding an anthocyanin transcription factor, wherein the transcription factor has a sequence comprising SEQ ID NO: 28 and said promoter comprises a sequence selected from the group consisting of SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46 and SEQ ID NO: 47. 
 
     
     
       9. The plant or plant part of  claim 8 , further comprising a nucleic acid sequence encoding a rate limiting anthocyanin pathway enzyme operably linked to a second promoter comprising a pathogen inducible regulatory element, wherein said promoter is native to corn cells. 
     
     
       10. The plant or plant part of  claim 5 , comprising a first cisgenic construct comprising a first pathogen inducible regulatory element operably linked to a nucleic acid sequence encoding a first signaling moiety and a second cisgenic construct comprising a pathogen inducible regulator element operably linked to a nucleic acid sequence encoding a second signaling moiety. 
     
     
       11. The plant or plant part of  claim 10  wherein the first and second pathogen inducible regulatory elements respond to different inducing agents. 
     
     
       12. The plant or plant part of  claim 5 , wherein said anthocyanin pathway factor is an enzyme selected from the group consisting of chalcone synthase, chalcone flavanone isomerase1, chalcone flavanone isomerase2, chalcone flavanone isomerase3, chalcone flavanone isomerase4, chalcone flavanone isomerase5, flavanone 3β-hydroxylase1, flavanone 3β-hydroxylase2, flavonoid 3′-hydroxylase, dihydroflavonol reductase1, dihydroflavonol 4-reductase, anthocyanin synthase, UDP-glucose flavonoid 3-O-glycosyltransferase, isoflavonoid synthase, flavonol synthase1, flavonol synthase2, cncr2 (cinnamoyl CoA reductase2), CCR4, dihydroflavonol-4-reductase, Flavonol synthase-like protein, NADPH dihydroflavonol reductase, Leucoanthocyanidin dioxygenase (LDOX), anthocyanidin synthase (ANS), flavanone 4-reductase, anthocyanidin 3-O-glucosyltransferase, glutathione S-transferase, anthocyanin acyltransferasel, and pale aleurone color1. 
     
     
       13. The plant or plant cell of  claim 10  comprising a first pathogen inducible regulatory element operably linked to a nucleic acid sequence encoding a transcription factor that enhances production of anthocyanins, and a second pathogen inducible regulator element operably linked to a nucleic acid sequence encoding a rate limiting anthocyanin pathway enzyme. 
     
     
       14. The plant of  claim 4 , wherein said plant is a corn plant and said cisgenic construct of the modified plant cells comprise a promoter sequence selected from the group consisting of SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46 and SEQ ID NO: 47. 
     
     
       15. A system for determining when to apply fungicide or insecticide treatments to a field comprising a plurality of plants, said system comprising:
 one or more sentry plants, wherein said sentry plants comprise the cisgenic construct of  claim 1 ; and 
 a detection system comprising a remote device for monitoring said plants to detect the expression of a detectable marker associated with said signaling moiety in said sentry plants. 
 
     
     
       16. The system of  claim 15  wherein said stress inducible regulatory element is a pathogen inducible regulatory element. 
     
     
       17. The system of  claim 16  wherein said signaling moiety is an anthocyanin pathway factor and the detection system detects changes in color in the cells of said sentry plants. 
     
     
       18. The system of  claim 17  wherein said detection system further comprises a computer having software, and said computer being configured to communicate with said remote device to receive data from the remote device and analyze said data to detect changes in color in said sentry plants resulting from an induction of said pathogen inducible promoter. 
     
     
       19. The system of  claim 18 , wherein the remote device is a camera that captures visual images, wherein the camera is
 i) mounted on a stationary pole; or 
 ii) mounted on an unmanned vehicle; or 
 iii) is fixed onto a drone or satellite. 
 
     
     
       20. The system of  claim 15  wherein said sentry plants further comprise a nucleic acid sequence encoding a rate limiting anthocyanin pathway enzyme operably linked to a second pathogen inducible regulatory element, optionally wherein said rate limiting anthocyanin pathway enzyme is chalcone synthase. 
     
     
       21. The system of  claim 19  wherein said detection system comprises
 a wireless controller including a processor; 
 a memory storing a program and a communication unit; and 
 a remote device configured to detect color changes in said plants and to communicate with the wireless controller, wherein the program, when executed by the processor, analyzes data received from the remote device and produce a signal when the data indicates the presence of plants with an altered change in color and optionally, the location of said plants with an altered change in color. 
 
     
     
       22. A method of treating pathogen-infected plants, said method comprising the steps of:
 planting a sentry plant in a field, wherein said sentry plant comprises a plurality of the modified plant cells according to  claim 1 ; 
 planting plants lacking said cisgenic construct adjacent to said sentry plant; 
 monitoring said field comprising said sentry plant for detection of said signaling moiety relative to said plants lacking said cisgenic construct; 
 applying an anti-pathogen treatment to said field in response to a detected alteration in expression of a detectable marker associated with said signaling moiety in said sentry plants relative to said plants lacking said cisgenic construct. 
 
     
     
       23. The method of  claim 22  wherein said signaling moiety is an anthocyanin pathway factor and the detectable marker is an alteration in color, and -the anti-pathogen treatment comprises an anti-fungal or insecticidal agent. 
     
     
       24. The method of  claim 23 , wherein the percentage of said sentry plants relative to the total plants in said field is less than about 25%, optionally wherein said sentry plants are interspersed with plants having a similar genetic background but lacking said cisgenic construct.